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Tissue-specific gene manipulations are widely used in genetically engineered mouse models. A single recombinase system, such as the one using Alb-Cre, has been commonly used for liver-specific genetic manipulations. However, most diseases are complex, involving multiple genetic changes and various cell types. A dual recombinase system is required for conditionally modifying different genes sequentially in the same cell or inducing genetic changes in different cell types within the same organism. A FlpO cDNA was inserted between the last exon and 3′-UTR of the mouse albumin gene in a bacterial artificial chromosome (BAC-Alb-FlpO). The founders were crossed with various reporter mice to examine the efficiency of recombination. Liver cancer tumorigenesis was investigated by crossing the FlpO mice with FSF-KrasG12D mice and p53frt mice (KPF mice). BAC-Alb-FlpO mice exhibited highly efficient recombination capability in both hepatocytes and intrahepatic cholangiocytes. No recombination was observed in the duodenum and pancreatic cells. BAC-Alb-FlpO-mediated liver-specific expression of mutant KrasG12D and conditional deletion of p53 gene caused the development of liver cancer. Remarkably, liver cancer in these KPF mice manifested a distinctive mixed hepatocellular carcinoma and cholangiocarcinoma phenotype. A highly efficient and liver-specific BAC-Alb-FlpO mouse model was developed. In combination with other Cre lines, different genes can be manipulated sequentially in the same cell, or distinct genetic changes can be induced in different cell types of the same organism. NEW & NOTEWORTHY: A liver-specific Alb-FlpO mouse line was generated. By coupling it with other existing CreERT or Cre lines, the dual recombinase approach can enable sequential gene modifications within the same cell or across various cell types in an organism for liver research through temporal and spatial gene manipulations. [ABSTRACT FROM AUTHOR]

Phylogenetic Trees are critical in human genome research for investigating human evolution and identifying disease-associated genetic markers. New high-throughput genome sequencing technologies raise an urgent need to develop statistical methods that can construct phylogenetic trees from long genome sequences with quick computation speeds, while considering various biological complexities. Though an ancestral mixture model has been proposed [Chen SC, Lindsay BG. Building mixture trees from binary sequence data. Biometrika. 2006;93(4):843–860. doi: 10.1093/biomet/93.4.843] to this end by allowing genetic mutations over generations, another essential evolution factor, genetic recombination, is missed. Therefore, in this paper, we develop a novel genetic recombination model for tree construction and propose to use Markov chain composite likelihood (MCCL) to make model estimation computationally feasible. To further reduce computation complexity, a hierarchical estimator is constructed to estimate unknown ancestral distributions through MCCL. Simulation studies and real data example show that our proposed methods perform well and fast, so have the potential for implementation in long sequence genome data. [ABSTRACT FROM AUTHOR]

Objectives To characterize VIM-type metallo-β-lactamase (MBL)-encoding genomic islands (GIs) in Pseudomonas aeruginosa and P. putida group isolates from Polish hospitals from 2001–2015/16. Methods Twelve P. aeruginosa and 20 P. putida group isolates producing VIM-like MBLs were selected from a large collection of these based on epidemiological and typing data. The organisms represented all major epidemic genotypes of these species spread in Poland with chromosomally located bla VIM gene-carrying integrons. The previously determined short-read sequences were complemented by long-read sequencing in this study. The comparative structural analysis of the GIs used a variety of bioinformatic tools. Results Thirty different GIs with bla VIM integrons were identified in the 32 isolates, of which 24 GIs from 26 isolates were integrative and conjugative elements (ICEs) of the clc family. These in turn were dominated by 21 variants of the GI2/ICE 6441 subfamily with a total of 19 VIM integrons, each inserted in the same position within the ICE's Tn 21 -like transposon Tn 4380. The three other ICEs formed a novel ICE 6705 subfamily, lacking Tn 4380 and having different VIM integrons located in another site of the elements. The remaining six non-ICE GIs represented miscellaneous structures. The presence of various integrons in the same ICE sublineage, and of the same integron in different GIs, indicated circulation and recombination of the integron-carrying genetic platforms across Pseudomonas species/genotypes. Conclusions Despite the general diversity of the bla VIM-carrying GIs in Pseudomonas spp. in Poland, a clear predominance of broadly spread and rapidly evolving clc -type ICEs was documented, confirming their significant role in antimicrobial resistance epidemiology. [ABSTRACT FROM AUTHOR]

Although germline BRCA mutations have been associated with adverse outcomes in prostate cancer (PC), understanding of the association between somatic/germline alterations in homologous recombination repair (HRR) genes and treatment outcomes in metastatic castration-resistant PC (mCRPC) is limited. The aim of this study was to investigate the prevalence and outcomes associated with somatic/germline HRR alterations, particularly BRCA1/2 , in patients initiating first-line (1L) mCRPC treatment with androgen receptor signalling inhibitors (ARSi) or taxanes. Data from 729 mCRPC patients were pooled for CAPTURE from four multicentre observational studies. Eligibility required 1L treatment with ARSi or taxanes, adequate tumour samples and biomarker panel results. Patients underwent paired normal and tumour DNA analyses by next-generation sequencing using a custom gene panel including ATM , BRCA1 , BRCA2 , BRIP1 , CDK12 , CHEK2 , FANCA , HDAC2 , PALB2 , RAD51B and RAD54L. Patients were divided into subgroups based on somatic/germline alteration(s): with BRCA1/2 mutations (BRCA); with HRR mutations except BRCA1/2 (HRR non-BRCA); and without HRR alterations (non-HRR). Patients without BRCA1/2 mutations were classified as non-BRCA. Radiographic progression-free survival (rPFS), progression-free survival 2 (PFS2) and overall survival (OS) were assessed. Of 729 patients, 96 (13.2%), 127 (17.4%) and 506 (69.4%) were in the BRCA, HRR non-BRCA and non-HRR subgroups, respectively. BRCA patients performed significantly worse for all outcomes than non-HRR or non-BRCA patients (P < 0.05), while PFS2 and OS were significantly shorter for BRCA than HRR non-BRCA patients (P < 0.05). HRR non-BRCA patients also had significantly worse rPFS, PFS2 and OS than non-HRR patients. Exploratory analyses suggested that for BRCA patients, there were no significant differences in outcomes associated with 1L treatment choice (ARSi or taxanes) or with the somatic/germline origin of the alterations. Worse outcomes were observed for mCRPC patients in the BRCA subgroup compared with non-BRCA subgroups, either HRR non-BRCA or non-HRR. Despite its heterogeneity, the HRR non-BRCA subgroup presented worse outcomes than the non-HRR subgroup. Screening early for HRR mutations, especially BRCA1/2 , is crucial in improving mCRPC patient prognosis. [Display omitted] • Almost one-third of mCRPC patients had ≥1 HRR alterations; about 13% had alterations in BRCA1/2 genes. • Patients with BRCA1/2 alterations, irrespective of somatic/germline origin, had worse outcomes than those without. • Outcomes for patients with BRCA1/2 alterations did not significantly differ by 1L treatment choice (ARSi or taxanes). • Identification of somatic/germline HRR alterations, namely BRCA1/2 , is key for targeted treatment and improved prognosis. [ABSTRACT FROM AUTHOR]

Echovirus 11 (E11) has gained attention owing to its association with severe neonatal infections. Due to the limited data available, the World Health Organization (WHO) considers public health risk to the general population to be low. The present study investigated the genetic variation and molecular evolution of E11 genomes collected from May to December 2023. Whole genome sequencing (WGS) was performed for 16 E11 strains. Phylogenetic analysis on WG showed how all Italian strains belonged to genogroup D5, similarly to other E11 strains recently reported in France and Germany all together aggregated into separate clusters. A cluster‐specific recombination pattern was also identified using phylogenetic analysis of different genome regions. Echovirus 6 was identified as the major recombinant virus in 3Cpro and 3Dpol regions. The molecular clock analysis revealed that the recombination event probably occurred in June 2018 (95% HPD interval: Jan 2016–Jan 2020). Shannon entropy analyses, within P1 region, showed how 11 amino acids exhibited relatively high entropy. Five of them were exposed on the canyon region which is responsible for receptor binding with the neonatal Fc receptor. The present study showed the recombinant origin of a new lineage of E11 associated with severe neonatal infections. [ABSTRACT FROM AUTHOR]

Modified vaccinia virus Ankara (MVA) has been widely tested in clinical trials as recombinant vector vaccine against infectious diseases and cancers in humans and animals. However, one biosafety concern about the use of MVA vectored vaccine is the potential forMVA to recombinewith naturally occurring orthopoxviruses in cells and hosts in which itmultiplies poorly and, therefore, producing viruses with mosaic genomes with altered genetic and phenotypic properties. We previously conducted co-infection and superinfection experiments with MVA vectored influenza vaccine (MVA-HANP) and a feline Cowpox virus (CPXV-No-F1) in Vero cells (that were semipermissive to MVA infection) and showed that recombination occurred in both coinfected and superinfected cells. In this study, we selected the putative recombinant viruses and performed genomic characterization of these viruses. Some putative recombinant viruses displayed plaque morphology distinct of that of the parental viruses. Our analysis demonstrated that they had mosaic genomes of different lengths. The recombinant viruses, with a genome more similar to MVA-HANP (>50%), rescued deleted and/or fragmented genes in MVA and gained new host ranges genes. Our analysis also revealed that someMVA-HANP contained a partially deleted transgene expression cassette and one recombinant virus contained part of the transgene expression cassette similar to that incomplete MVA-HANP. The recombination in co-infected and superinfected Vero cells resulted in recombinant viruses with unpredictable biological and genetic properties as well as recovery of delete/fragmented genes in MVA and transfer of the transgene into replication competent CPXV. These results are relevant to hazard characterization and risk assessment of MVA vectored biologicals. [ABSTRACT FROM AUTHOR]

Aims: Diabetic osteoporosis (DOP) is the most common secondary form of osteoporosis. Diabetes mellitus affects bone metabolism; however, the underlying pathophysiological mechanisms remain unclear. Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) expression is upregulated in conditions characterized by vascular injury, such as atherosclerosis, hypertension, and diabetes. Additionally, Notch, HIF-1α, and VEGF are involved in angiogenesis and bone formation. Therefore, we aimed to investigate the expression of Notch, HIF-1α, and VEGF in the LOX-1 silencing state. Methods: Rat bone H-type vascular endothelial cells (THVECs) were isolated and cultured in vitro. Cell identification was performed using immunofluorescent co-expression of CD31 and Emcn. Lentiviral silencing vector (LV-LOX-1) targeting LOX-1 was constructed using genetic recombination technology and transfected into the cells. The experimental groups included the following: NC group, HG group, LV-LOX-1 group, LV-CON group, HG + LV-LOX-1 group, HG + LV-CON group, HG + LV-LOX-1 + FLI-06 group, HG + LV-CON + FLI-06 group, HG + LV-LOX-1 + LW6 group, and HG + LV-CON + LW6 group. The levels of LOX-1, Notch, Hif-1α, and VEGF were detected using PCR and WB techniques to investigate whether the expression of LOX-1 under high glucose conditions has a regulatory effect on downstream molecules at the gene and protein levels, as well as the specific molecular mechanisms involved. Results: High glucose (HG) conditions led to a significant increase in LOX-1 expression, leading to inhibition of angiogenesis, whereas silencing LOX-1 can reverse this phenomenon. Further analysis reveals that changes in LOX-1 will promote changes in Notch/HIF-1α and VEGF. Moreover, Notch mediates the activation of HIF-1α and VEGF. Conclusions: The activation of LOX-1 and the inhibition of Notch/HIF-1α/VEGF in THVECs are the main causes of DOP. These findings contribute to our understanding of the pathogenesis of DOP and offer a novel approach for preventing and treating osteoporosis. [ABSTRACT FROM AUTHOR]

Although the defining events of meiosis are highly conserved, such as genetic recombination and ploidy reduction, the regulation of meiotic entry and divisions is distinct between species and sexes. In mammals, the complex cellular milieu dictates the developmental timing and duration of meiosis and hinges on complex signaling from somatic cells. The unique and dramatic chromosomal events of meiosis and gametogenesis require diverse mechanisms of regulation of gene expression and cell cycle progression. The functional study of meiotic regulatory elements in animals is markedly hampered by the inability to efficiently recapitulate the entire process in cell culture paradigms. However, increasingly powerful genomic analyses are illuminating key aspects of meiotic gene expression such as transcription control, transcriptome epigenetics, and post-transcriptional regulation. Meiosis is essential for gamete production in all sexually reproducing organisms. It entails two successive cell divisions without DNA replication, producing haploid cells from diploid ones. This process involves complex morphological and molecular differentiation that varies across species and between sexes. Specialized genomic events like meiotic recombination and chromosome segregation are tightly regulated, including preparation for post-meiotic development. Research in model organisms, notably yeast, has shed light on the genetic and molecular aspects of meiosis and its regulation. Although mammalian meiosis research faces challenges, particularly in replicating gametogenesis in vitro , advances in genetic and genomic technologies are providing mechanistic insights. Here we review the genetics and molecular biology of meiotic gene expression control, focusing on mammals. [ABSTRACT FROM AUTHOR]

Human adenovirus (HAdV) is a significant viral pathogen causing severe acute respiratory infections (SARIs) in children. To improve the understanding of type distribution and viral genetic characterization of HAdV in severe cases, this study enrolled 3404 pediatric SARI cases from eight provinces of China spanning 2017–2021, resulting in the acquisition of 112 HAdV strains. HAdV‐type identification, based on three target genes (penton base, hexon, and fiber), confirmed the diversity of HAdV types in SARI cases. Twelve types were identified, including species B (HAdV‐3, 7, 55), species C (HAdV‐1, 2, 6, 89, 108, P89H5F5, Px1/Ps3H1F1, Px1/Ps3H5F5), and E (HAdV‐4). Among these, HAdV‐3 exhibited the highest detection rate (44.6%), followed by HAdV‐7 (19.6%), HAdV‐1 (12.5%), and HAdV‐108 (9.8%). All HAdV‐3, 7, 55, 4 in this study belonged to dominant lineages circulating worldwide, and the sequences of the three genes demonstrated significant conservation and stability. Concerning HAdV‐C, excluding the novel type Px1/Ps3H1F1 found in this study, the other seven types were detected both in China and abroad, with HAdV‐1 and HAdV‐108 considered the two main types of HAdV‐C prevalent in China. Two recombinant strains, including P89H5F5 and Px1/Ps3H1F1, could cause SARI as a single pathogen, warranting close monitoring and investigation for potential public health implications. In conclusion, 5 years of SARI surveillance in China provided crucial insights into HAdV‐associated respiratory infections among hospitalized pediatric patients. [ABSTRACT FROM AUTHOR]

Background: Porcine circovirus type 2 (PCV2) is a globally prevalent and recurrent pathogen that primarily causes slow growth and immunosuppression in pigs. Porcine circovirus type 3 (PCV3), a recently discovered virus, commonly leads to reproductive disorders in pigs and has been extensively disseminated worldwide. Infectionwith a single PCV subtype alone does not induce severe porcine circovirusassociated diseases (PCVD), whereas concurrent co-infection with PCV2 and PCV3 exacerbates the clinical manifestations. Pseudorabies (PR), a highly contagious disease in pigs, pose a significant threat to the swine industry in China. Methods: In this study, recombinant strains named rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 was constructed by using a variant strain XJ of pseudorabies virus (PRV) as the parental strain, with the TK/gE/gI genes deleted and simultaneous expression of PCV2 Cap, PCV3 Cap, and IL-4. The two recombinant strains obtained by CRISPR/Cas gE gene editing technology and homologous recombination technology has genetic stability in baby hamster Syrian kidney-21 (BHK-21) cells and is safe to mice. Results: rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 exhibited good safety and immunogenicity in mice, inducing high levels of antibodies, demonstrated 100% protection against the PRV challenge in mice, reduced viral loads and mitigated pathological changes in the heart, lungs, spleen, and lymph nodes during PCV2 challenge. Moreover, the recombinant viruses with the addition of IL-4 as a molecular adjuvant outperformed the non-addition group in most indicators. Conclusion: rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 hold promise as recombinant vaccines for the simultaneous prevention of PCV2, PCV3, and PRV, while IL-4, as a vaccine molecular adjuvant, effectively enhances the immune response of the vaccine. [ABSTRACT FROM AUTHOR]

Since 2013, the porcine reproductive and respiratory syndrome virus type 2 (PRRSV-2), lineage 1.8 (NADC30-like PRRSV) has emerged and become widely prevalent in China. The NADC30-like PRRSV poses significant challenges for disease control, primarily because of its propensity for frequent mutations and recombinations. We successfully isolated and identified a NADC30-like strain, designated SCCD22, in Chengdu, Sichuan Province, China. We meticulously examined the genetic recombination properties and evaluated its pathogenicity in 28-day-old piglets. SCCD22 showed 93.02% nucleotide homology with the NADC30 PRRSV strain, and its non-structural protein 2 coding region showed the same 131 amino acid deletion pattern as that seen in NADC30. Furthermore, we identified two recombination events in SCCD22: one in the NSP2 region (1,028-3,290nt), where it was highly similar to the JXA1-like strain GZ106; and another in the NSP10~ 12 region (9,985-12,279nt), closely resembling the NADC30-like strain CY2-1604. Piglets infected with SCCD22 exhibited clinical symptoms such as elevated body temperature, prolonged fever, reduced appetite, and roughened fur. Postmortem examinations underscored the typical lung pathology associated with PRRSV, indicating that the lungs were the primary affected organs. Furthermore, extended viral shedding accompanied by progressive viremia was observed in the serum and nasal excretions of infected piglets. In summary, this study reports a domestic PRRSV recombination strain in the Sichuan Province that can provide critical insights into preventing and controlling PRRSV in this region. [ABSTRACT FROM AUTHOR]

Bovine parainfluenza virus type 3 (BPIV-3) is one of the major pathogens of the bovine respiratory disease complex (BRDC). BPIV-3 surveillance in China has been quite limited. In this study, we used PCR to test 302 cattle in China, and found that the positive rate was 4.64% and the herd-level positive rate was 13.16%. Six BPIV-3C strains were isolated and confirmed by electron microscopy, and their titers were determined. Three were sequenced by next-generation sequencing (NGS). Phylogenetic analyses showed that all isolates were most closely related to strain NX49 from Ningxia; the genetic diversity of genotype C strains was lower than strains of genotypes A and B; the HN, P, and N genes were more suitable for genotyping and evolutionary analyses of BPIV-3. Protein variation analyses showed that all isolates had mutations at amino acid sites in the proteins HN, M, F, and L. Genetic recombination analyses provided evidence for homologous recombination of BPIV-3 of bovine origin. The virulence experiment indicated that strain Hubei-03 had the highest pathogenicity and could be used as a vaccine candidate. These findings apply an important basis for the precise control of BPIV-3 in China. [ABSTRACT FROM AUTHOR]

Chromosomal inversions may play a central role in speciation given their ability to locally reduce recombination and therefore genetic exchange between diverging populations. We analyzed long- and short-read whole-genome data from sympatric and allopatric populations of 2 Drosophila virilis group species, Drosophila montana and Drosophila flavomontana , to understand if inversions have contributed to their divergence. We identified 3 large alternatively fixed inversions on the X chromosome and one on each of the autosomes 4 and 5. A comparison of demographic models estimated for inverted and noninverted (colinear) chromosomal regions suggests that these inversions arose before the time of the species split. We detected a low rate of interspecific gene flow (introgression) from D. montana to D. flavomontana , which was further reduced inside inversions and was lower in allopatric than in sympatric populations. Together, these results suggest that the inversions were already present in the common ancestral population and that gene exchange between the sister taxa was reduced within inversions both before and after the onset of species divergence. Such ancestrally polymorphic inversions may foster speciation by allowing the accumulation of genetic divergence in loci involved in adaptation and reproductive isolation inside inversions early in the speciation process, while gene exchange at colinear regions continues until the evolving reproductive barriers complete speciation. The overlapping X inversions are particularly good candidates for driving the speciation process of D. montana and D. flavomontana , since they harbor strong genetic incompatibilities that were detected in a recent study of experimental introgression. [ABSTRACT FROM AUTHOR]

During meiosis, genetic recombination is initiated by the formation of many DNA double-strand breaks (DSBs) catalysed by the evolutionarily conserved topoisomerase-like enzyme, Spo11, in preferred genomic sites known as hotspots. DSB formation activates the Tel1/ATM DNA damage responsive (DDR) kinase, locally inhibiting Spo11 activity in adjacent hotspots via a process known as DSB interference. Intriguingly, in S. cerevisiae, over short genomic distances (<15 kb), Spo11 activity displays characteristics of concerted activity or clustering, wherein the frequency of DSB formation in adjacent hotspots is greater than expected by chance. We have proposed that clustering is caused by a limited number of sub-chromosomal domains becoming primed for DSB formation. Here, we provide evidence that DSB clustering is abolished when meiotic prophase timing is extended via deletion of the NDT80 transcription factor. We propose that extension of meiotic prophase enables most cells, and therefore most chromosomal domains within them, to reach an equilibrium state of similar Spo11-DSB potential, reducing the impact that priming has on estimates of coincident DSB formation. Consistent with this view, when Tel1 is absent but Ndt80 is present and thus cells are able to rapidly exit meiotic prophase, genome-wide maps of Spo11-DSB formation are skewed towards pericentromeric regions and regions that load pro-DSB factors early—revealing regions of preferential priming—but this effect is abolished when NDT80 is deleted. Our work highlights how the stochastic nature of Spo11-DSB formation in individual cells within the limited temporal window of meiotic prophase can cause localised DSB clustering—a phenomenon that is exacerbated in tel1Δ cells due to the dual roles that Tel1 has in DSB interference and meiotic prophase checkpoint control. Author summary: Genetic variation arises in sexually reproducing organisms via the combination of two processes: outbreeding and meiosis. Outbreeding ensures that individuals are unique composites of their genetically distinct parents, whereas meiosis—a specialised form of cell division—creates genetic variation within the gametes used during breeding (eggs and sperm in humans). During the early stages of meiosis—termed prophase—hundreds of DNA breaks are generated within parental chromosomes. Repair of these DNA breaks generates novel combinations of gene types (alleles). Despite many steps of meiosis being evolutionarily conserved from yeast to humans, it is poorly understood how DNA break formation is regulated to ensure that DNA breaks are evenly spread across all chromosomes in order to generate genetic diversity. Here, we utilise the model eukaryote, Saccharomyces cerevisiae, (budding yeast) to investigate the interplay between a key protein involved in DNA break recognition (Tel1, the yeast version of the mammalian gene, ATM), and the length of meiotic prophase controlled by the transcription factor Ndt80. Our observations indicate that the clustering of DNA breaks that happens when Tel1 is absent is suppressed by extending the time window of meiotic prophase, providing novel insight into the regulation of genetic variation within sexually reproducing organisms. [ABSTRACT FROM AUTHOR]

The large-fruited fresh-market tomato cultivated in the U.S. represents a unique fruit market class of contemporary (modern) tomatoes for direct consumption. The genomes of F2 plants from crosses between inbred contemporary U.S. large-fruited fresh-market tomatoes were sequenced. 516 F2 individual plants randomly selected from five different biparental segregating populations were used for DNA extraction. The polymerase chain reaction (PCR)-free, paired-end (2 × 150 bp) sequencing libraries (350 bp DNA fragment length) were prepared, and sequenced on average 5 Gb for each plant using the Illumina next-generation sequencing technologies [1,2]. Raw Illumina reads with adapter contamination and/or uncertain nucleotides constitute (Ns, >10 % of either read; Q-score 5 or lower, >50 % of either read) were removed. This data article will contribute to improving our knowledge of the genetic recombination and variation in tomato.

ABSTRACT The objective of this study was to characterize a novel circulating recombinant form of human immunodeficiency virus type 1 (HIV-1) among people living with HIV in Karachi, Pakistan. We conducted near-full-length genome (NFLG) sequencing on eight samples exhibiting D/G recombination signals in the pol gene region. We successfully obtained NFLG sequences (790–9,614; with reference to the HXB2 genome) from four of the eight samples and then conducted phylogenetic and recombination analyses on them. The four NFLG sequences from our study and one DG unique recombinant form previously identified in the United Kingdom (GenBank accession: MF109700) formed a distinct monophyletic cluster with an Shimodaira-Hasegawa approximate likelihood ratio test node support value of 100%. Bootscan analyses of the five NFLG sequences of DG recombinants showed that all five NFLGs shared the same unique mosaic pattern of recombination breakpoints between D and G clades, with two D fragments in the pol and vif regions inserted into a G backbone. Subregion phylogenetic analyses confirmed these sequences to be a novel circulating recombinant form (CRF) composed of subtypes D and G. The DG recombinant sequences were eventually designated as CRF152_DG by the Los Alamos HIV Sequence Database staff.IMPORTANCEIn Pakistan, the genetic diversity of human immunodeficiency virus type 1 (HIV-1) is becoming increasingly complex, compared to the early years of the epidemic that started after the detection of the first cases of HIV-1 in 1987 in Karachi. Based on the available molecular studies, two dominant HIV-1 clades, sub-subtype A1 and CRF02_AG, have been found to co-circulate with other clades, namely B, C, D, G, CRF01_AE, CRF35_A1D, and CRF56_cpx, in various urban areas of Pakistan. Several novel recombinant forms have also been detected. This first report of CRF152_DG highlights the complex nature of the HIV epidemic in Pakistan and emphasizes the importance of continual molecular surveillance (ideally based on whole-genome sequences) of HIV.

Hand, foot, and mouth disease (HFMD) is one of the most common class C infectious diseases, posing a serious threat to public health worldwide. Enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16) have been regarded as the major pathogenic agents of HFMD; however, since an outbreak caused by coxsackievirus A6 (CV-A6) in France in 2008, CV-A6 has gradually become the predominant pathogen in many regions. CV-A6 infects not only children but also adults, and causes atypical clinical symptoms such as a more generalized rash, eczema herpeticum, high fever, and onychomadesis, which are different from the symptoms associated with EV-A71 and CV-A16. Importantly, the rate of genetic recombination of CV-A6 is high, which can lead to changes in virulence and the rapid evolution of other characteristics, thus posing a serious threat to public health. To date, no specific vaccines or therapeutics have been approved for CV-A6 prevention or treatment, hence it is essential to fully understand the relationship between recombination and evolution of this virus. Here, we systematically review the genetic recombination events of CV-A6 that have occurred worldwide and explore how these events have promoted virus evolution, thus providing important information regarding future HFMD surveillance and prevention.

Abstract Streptococcus suis is a zoonotic pathogen that causes a major health problem in the pig production industry worldwide. Spain is one of the largest pig producers in the world. This work aimed to investigate the genetic and phenotypic features of invasive S. suis isolates recovered in Spain. A panel of 156 clinical isolates recovered from 13 Autonomous Communities, representing the major pig producers, were analysed. MLST and serotyping analysis revealed that most isolates (61.6%) were assigned to ST1 (26.3%), ST123 (18.6%), ST29 (9.6%), and ST3 (7.1%). Interestingly, 34 new STs were identified, indicating the emergence of novel genetic lineages. Serotypes 9 (27.6%) and 1 (21.8%) prevailed, followed by serotypes 7 (12.8%) and 2 (12.2%). Analysis of 13 virulence-associated genes showed significant associations between ST, serotype, virulence patterns, and clinical features, evidencing particular virulence traits associated with genetic clusters. The pangenome was generated, and the core genome was distributed in 7 Bayesian groups where each group included a variable set of over- and under-represented genes of different categories. The study provides comprehensive data and knowledge to improve the design of new vaccines, antimicrobial treatments, and bacterial typing approaches.

The article focuses on the development and utility of the BAC-Alb-FlpO transgenic mouse line as a novel tool for liver disease research. Topics include its ability to express FlpO recombinase for high-efficiency genetic manipulation, its application in generating liver cancer models, and its potential combination with existing recombination systems for advanced studies on liver diseases.

The transition from text to images as the primary form of information transmission has recently increased the need for secure and effective encryption techniques due to the expanding information dimensions. The color picture encryption algorithm utilizing chaotic mapping is limited by a small chaotic range, unstable chaotic state, and lengthy encryption duration. This study integrates the Ackley function and the Styblinski–Tang function into a novel two-dimensional hyperchaotic map for optimization testing. A randomness test is run on the chaotic sequence created by the system to check that the new chaotic system can better sustain the chaotic state. This study introduces two techniques, genetic recombination and clock diffusion, to simultaneously disperse and mix images at the bit level. This study utilizes chaotic sequences in genetic recombination and clock drift to propose an image encryption technique. The data indicates that the method demonstrates high encryption efficiency. At the same time, the key also successfully passed the NIST randomness test, verifying its sensitivity and randomness. The algorithm’s dependability has been demonstrated and can be utilized for color image encryption.

During the routine surveillance, we isolated nine H4N6 subtype avian influenza viruses (AIVs) in Jiangsu Province, China, in March 2023. Phylogenetic analysis revealed that nine H4N6 viruses belonged to the Eurasian lineage and underwent complex genetic recombination among Asian countries during their evolution. It is particularly noteworthy that the PB2 and PB1 genes of our representative virus were descended from clade 2.3.4.4b H5 high-pathogenic AIVs in Japan. Mutations of D3V and D622G in PB1, N66S in PB1-F2, N30D, I43M, and T215A in M1, and P42S and I106M in NS1 were observed in nine isolates, which may increase the pathogenicity of the viruses in mice. The receptor binding analysis showed that the tested H4N6 virus could bind to both avian-type and human-type receptors. Vitro infection kinetics revealed that the representative virus could efficiently replicate in mammalian cells, including MDCK and 293T cells. Pathogenicity tests in mice indicated that the representative virus could replicate in nasal turbinates and lungs without prior adaptation. Our data reveal the potential public health issues represented by H4N6 viruses from wild birds and highlight the need to strengthen routine surveillance of wild birds. [ABSTRACT FROM AUTHOR]

Environmental stress at high altitudes drives the development of distinct adaptive mechanisms in plants. However, studies exploring the genetic adaptive mechanisms of high-altitude plant species are scarce. In the present study, we explored the high-altitude adaptive mechanisms of plants in the Himalayas through whole-genome resequencing. We studied two widespread members of the Himalayan endemic alpine genus Roscoea (Zingiberaceae): R. alpina (a selfing species) and R. purpurea (an outcrossing species). These species are distributed widely in the Himalayas with distinct non-overlapping altitude distributions; R. alpina is distributed at higher elevations, and R. purpurea occurs at lower elevations. Compared to R. purpurea, R. alpina exhibited higher levels of linkage disequilibrium, Tajima's D, and inbreeding coefficient, as well as lower recombination rates and genetic diversity. Approximately 96.3% of the genes in the reference genome underwent significant genetic divergence (FST ≥ 0.25). We reported 58 completely divergent genes (FST = 1), of which only 17 genes were annotated with specific functions. The functions of these genes were primarily related to adapting to the specific characteristics of high-altitude environments. Our findings provide novel insights into how evolutionary innovations promote the adaptation of mountain alpine species to high altitudes and harsh habitats. [ABSTRACT FROM AUTHOR]

Aiming at the simultaneous scheduling problem of scheduling jobs and automated guide vehicles (AGVs) in the flexible workshop system, consider building the objective function to minimize maximum processing machine duration, single AGV handling time, and total AGV handling time in the case of a finite number of AGVs and processing machines. Design an improved algorithm that combines NSGA-II (non-dominated sorting genetic algorithms) and clonal selection algorithm (CSA) to solve such problems. Firstly, the workpiece, the processing machine and the AGV are used for three-part coding. Secondly, the total score of non-dominated ranking and objective function value size sorting is introduced to stratify the population, so as to effectively retain excellent individuals. Thirdly, for the cloned population, different probabilities of variation are adopted for different levels of populations, and the genetic recombination of internal exchange and uniform cross-mixing exchange of chromosomes is carried out to effectively improve the diversity of the population and prevent it from falling into local optimum. Finally, three sets of comparative experiments verify that the algorithm has the advantages of short running time, high stability and good convergence when exploring the optimal solution. [ABSTRACT FROM AUTHOR]

DVGs (Defective Viral Genomes) are prevalent in RNA virus infections. In this investigation, we conducted an analysis of high-throughput sequencing data and observed widespread presence of DVGs in SARS-CoV-2. Comparative analysis between SARS-CoV-2 and diverse DNA viruses revealed heightened susceptibility to damage and increased sequencing sample heterogeneity within the SARS-CoV-2 genome. Whole-genome sequencing depth variability analysis exhibited a higher coefficient of variation for SARS-CoV-2, while DVG analysis indicated a significant proportion of recombination sites, signifying notable genome heterogeneity and suggesting that a large proportion of assembled virus particles contain incomplete RNA sequences. Moreover, our investigation explored the sequencing depth and DVG content differences among various strains. Our findings revealed that as the virus evolves, there is a notable increase in the proportion of intact genomes within virus particles, as evidenced by third-generation sequencing data. Specifically, the proportion of intact genome in the Omicron strain surpassed that of the Delta and Alpha strains. This observation effectively elucidates the heightened infectiousness of the Omicron strain compared to the Delta and Alpha strains. We also postulate that this improvement in completeness stems from enhanced virus assembly capacity, as the Omicron strain can promptly facilitate the binding of RNA and capsid protein, thereby reducing the exposure time of vulnerable virus RNA in the host environment and significantly mitigating its degradation. Finally, employing mathematical modeling, we simulated the impact of DVG effects under varying environmental factors on infection characteristics and population evolution. Our findings provide an explanation for the close association between symptom severity and the extent of virus invasion, as well as the substantial disparity in population infection characteristics caused by the same strain under distinct environmental conditions. This study presents a novel approach for future virus research and vaccine development. [ABSTRACT FROM AUTHOR]

During the investigations of discomycetes in Yunnan, China, five species of Tatraea were discovered on decayed, decorticated oak trees or unidentified wood. All species have typical disc-like, large fruiting bodies with grey, brown or greyish-green colors. The ITS sequence analysis showed that they belong to Tatraea (Helotiaceae, Helotiales) and the LSU and ITS combination revealed a different topology within the genus. Four species, T. clepsydriformis, T. griseoturcoisina, T. yunnanensis and T. yuxiensis were established as new species, and T. aseptata was collected and described on oak woods. The pairwise homoplasy index (PHI) test results indicated that there is no significant genetic recombination (Fw = 1.0) between all related species pairs. All the species described here are supported by descriptions, illustrations and multi-gene analyses. [ABSTRACT FROM AUTHOR]

Context. Researchers have been accumulating information concerning the locations of genes and quantitative trait loci (QTLs) in cultivated oat (Avena sativa L.) for more than 100 years. Aims. The aim of this work was to create an inventory of genes and QTLs found in cultivated hexaploid oat and produce tools to make this resource more useful. Methods. By using the positions of perfectly matched, single nucleotide polymorphism markers, each centimorgan (cM) location along the consensus map was assigned to a location on the OT3098 v2 physical map found on the GrainGenes database website (https://wheat.pw.usda.gov/jb/?data=/ggds/oat-ot3098v2-pepsico). This information was then used to assign physical locations to the genes and QTLs in the inventory, where possible. Key results. A table comparing the major genetic maps of hexaploid oats to each other, to the 2018 oat consensus map, and to physical chromosomes was produced. Genome browser tracks aligning the consensus map regions and the locations of the genes and QTLs to OT3098 v2 were added to GrainGenes. Conclusions. Many oat genes and QTLs identified using genetic mapping could be assigned positions on physical oat chromosomes. However, many of these assigned regions are quite long, owing to the presence of large areas of reduced recombination. Specific examples of identified patterns of recombination between the genetic and physical maps and validated gene and QTL locations are discussed. Implications. These resources will assist researchers performing comparative genetic and physical mapping in oat. [ABSTRACT FROM AUTHOR]

Feline leukemia virus (FeLV) is an exogenous retrovirus that causes malignant hematopoietic disorders in domestic cats, and its virulence may be closely associated with viral sequences. FeLV is classified into several subgroups, including A, B, C, D, E, and T, based on viral receptor interference properties or receptor usage. However, the transmission manner and disease specificity of the recombinant viruses FeLV-D and FeLV-B remain unclear. The aim of this study was to understand recombination events between exogenous and endogenous retroviruses within a host and elucidate the emergence and transmission of recombinant viruses. We observed multiple recombination events involving endogenous retroviruses (ERVs) in FeLV from a family of domestic cats kept in one house; two of these cats (ON-T and ON-C) presented with lymphoma and leukemia, respectively. Clonal integration of FeLV-D was observed in the ON-T case, suggesting an association with FeLV-D pathogenesis. Notably, the receptor usage of FeLV-B observed in ON-T was mediated by feline Pit1 and feline Pit2, whereas only feline Pit1 was used in ON-C. Furthermore, XR-FeLV, a recombinant FeLV containing an unrelated sequence referred to the X-region, which is homologous to a portion of the 5′-leader sequence of Felis catus endogenous gammaretrovirus 4 (FcERV-gamma4), was isolated. Genetic analysis suggested that most recombinant viruses occurred de novo; however, the possibility of FeLV-B transmission was also recognized in the family. This study demonstrated the occurrence of multiple recombination events between exogenous and endogenous retroviruses in domestic cats, highlighting the contribution of ERVs to pathogenic recombinant viruses. IMPORTANCE Feline leukemia virus subgroup A (FeLV-A) is primarily transmitted among cats. During viral transmission, genetic changes in the viral genome lead to the emergence of novel FeLV subgroups or variants with altered virulence. We isolated three FeLV subgroups (A, B, and D) and XR-FeLV from two cats and identified multiple recombination events in feline endogenous retroviruses (ERVs), such as enFeLV, ERV-DC, and FcERV-gamma4, which are present in the cat genome. This study highlights the pathogenic contribution of ERVs in the emergence of FeLV-B, FeLV-D, and XR-FeLV in a feline population. [ABSTRACT FROM AUTHOR]

South Korea has experienced outbreaks of foot-and-mouth disease (FMD) of serotypes O and A, leading to nationwide vaccination with a bivalent vaccine. Since the FMD virus (FMDV) Asia1 group-V genotype occurred in North Korea in 2007, an Asia1/MOG/05 vaccine strain belonging to the Asia1 group-V genotype was developed using a genetic recombination method (Asia1/MOG/05-R). This study aimed to evaluate the antigen productivity and viral inactivation kinetics of Asia1/MOG/05-R to assess its commercial viability. The antigen yield of Asia1/MOG/05-R produced in flasks and bioreactors was approximately 4.0 μg/mL. Binary ethylenimine (BEI) inactivation kinetics of Asia1/MOG/05-R showed that 2 mM and 1.0 mM BEI treatment at 26 °C and 37 °C, respectively, resulted in a virus titer <10−7 TCID50/mL within 24 h, meeting the inactivation kinetics criteria. During incubation at 26 °C and 37 °C, 10% antigen loss occurred, but not due to BEI treatment. When pigs were inoculated twice with the Asia1/MOG/05-R antigen, the virus neutralization titer increased to approximately 1:1000; therefore, it can sufficiently protect against Asia1/MOG/05-R and Asia1 Shamir viruses. The Asia1/MOG/05-R will be useful as a vaccine strain for domestic antigen banks. [ABSTRACT FROM AUTHOR]

Streptococcus suis is a zoonotic pathogen that causes a major health problem in the pig production industry worldwide. Spain is one of the largest pig producers in the world. This work aimed to investigate the genetic and phenotypic features of invasive S. suis isolates recovered in Spain. A panel of 156 clinical isolates recovered from 13 Autonomous Communities, representing the major pig producers, were analysed. MLST and serotyping analysis revealed that most isolates (61.6%) were assigned to ST1 (26.3%), ST123 (18.6%), ST29 (9.6%), and ST3 (7.1%). Interestingly, 34 new STs were identified, indicating the emergence of novel genetic lineages. Serotypes 9 (27.6%) and 1 (21.8%) prevailed, followed by serotypes 7 (12.8%) and 2 (12.2%). Analysis of 13 virulence-associated genes showed significant associations between ST, serotype, virulence patterns, and clinical features, evidencing particular virulence traits associated with genetic clusters. The pangenome was generated, and the core genome was distributed in 7 Bayesian groups where each group included a variable set of over- and under-represented genes of different categories. The study provides comprehensive data and knowledge to improve the design of new vaccines, antimicrobial treatments, and bacterial typing approaches. [ABSTRACT FROM AUTHOR]

Phylogenetic trees can be built from B cell receptor (BCR) sequences and used to study somatic hypermutation and selection. The unique biology of B cells has led to the development of B cell-specific phylogenetic methods. Single-cell sequencing can improve tree building with paired heavy and light chain sequences and (potentially) with associated transcriptomic information. Phylogenetic trees can be used to study B cell migration, differentiation, and evolution over time, but more work is needed to study these processes in a model-based manner. Phylogenetic trees built from B cell receptor (BCR) sequences can trace the history of mutation and selection in B cell clones. Advances in single-cell sequencing can significantly improve all aspects of B cell tree building. Further, there is potential for new methods using single-cell data to drive exciting advances in tracking cellular migration, differentiation, and evolution over time. The widespread availability of single-cell RNA sequencing (scRNA-seq) has led to the development of new methods for understanding immune responses. Single-cell transcriptome data can now be paired with B cell receptor (BCR) sequences. However, RNA from BCRs cannot be analyzed like most other genes because BCRs are genetically diverse within individuals. In humans, BCRs are shaped through recombination followed by mutation and selection for antigen binding. As these processes co-occur with cell division, B cells can be studied using phylogenetic trees representing the mutations within a clone. B cell trees can link experimental timepoints, tissues, or cellular subtypes. Here, we review the current state and potential of how B cell phylogenetics can be combined with single-cell data to understand immune responses. [ABSTRACT FROM AUTHOR]

False smut, caused by Villosiclava virens, is becoming increasingly serious in modern rice production systems, leading to yield losses and quality declines. Successful infection requires efficient acquisition of sucrose, abundant in rice panicles, as well as other sugars. Sugar transporters (STPs) may play an important role in this process. STPs belong to a major facilitator superfamily, which consists of large multigenic families necessary to partition sugars between fungal pathogens and their hosts. This study identified and characterized the STP family of V. viren, and further analyzed their gene functions to uncover their roles in interactions with rice. Through genome-wide and systematic bioinformatics analyses, 35 STPs were identified from V.virens and named from VvSTP1 to VvSTP35. Transmembrane domains, gene structures, and conserved motifs of VvSTPs have been identified and characterized through the bioinformatic analysis. In addition, a phylogenetic analysis revealed relationship between VvSTPs and STPs from the other three reference fungi. According to a qRT-PCR and RNA-sequencing analysis, VvSTP expression responded differently to different sole carbon sources and H2O2 treatments, and changed during the pathogenic process, suggesting that these proteins are involved in interactions with rice and potentially functional in pathogenesis. In total, 12 representative VvSTPs were knocked out through genetic recombination in order to analyze their roles in pathogenicity of V. virens. The knock-out mutants of VvSTPs showed little difference in mycelia growth and conidiation, indicating a single gene in this family cannot influence vegetative growth of V. virens. It is clear, however, that these mutants result in a change in infection efficiency in a different way, indicating that VvSTPs play an important role in the pathogenicity of virens. This study is expected to contribute to a better understanding of how host-derived sugars contribute to V. virens pathogenicity. [ABSTRACT FROM AUTHOR]

Insertional mutation, phenotypic evaluation, and mutated gene cloning are widely used to clone genes from scratch. Exogenous genes can be integrated into the genome during non-homologous end joining (NHEJ) of the double-strand breaks of DNA, causing insertional mutation. The random insertional mutant library constructed using this method has become a method of forward genetics for gene cloning. However, the establishment of a random insertional mutant library requires a high transformation efficiency of exogenous genes. Many microalgal species show a low transformation efficiency, making constructing random insertional mutant libraries difficult. In this study, we established a highly efficient transformation method for constructing a random insertional mutant library of Nannochloropsis oceanica, and tentatively tried to isolate its genes to prove the feasibility of the method. A gene that may control the growth rate and cell size was identified. This method will facilitate the genetic studies of N. oceanica, which should also be a reference for other microalgal species. [ABSTRACT FROM AUTHOR]

Background: The rapid rate of mutation of corona virus, which is a significant characteristic, suggests that as it moves across various habitats, the viral genome undergoes new alterations. The objective of this study was to analyze SARS-COV2 ORF8 gene sequence variation among patients in Erbil city. Materials & Methods: During the period of January to December 2022, ten SARS-CoV-2 positive patients (4 female and 6 male), who were detected during the pandemic at Erbil central Laboratory aged (20-70) years enrolled in this cross-sectional study. The single strand RNA extracted from patients and transformed to double stranded complementary DNA. The primers (forward and reveres)- were designed by Bioinformatics program, (Primer3 Plus). Polymerase chain reaction amplification for ORF8 gene was carried out. The products ran on gel electrophoresis to visualize the DNA products. Each species was bi-directionally sequenced to get sequence of DNA strand according to forward primer. The sequence editing was analyzed using BLAST NCBI to indicate the homology from closest species. Phylogenetic tree was constructed. Results: In comparing to the wild type that stated by NCBI (Genbank Reference accession number: OP732758.1) sequencing of our samples showed mutations at different position in which codon number 35 nucleotide A changed to C which results in replacement of an amino acid Aspartic Acid by Alanine, in addition to that in codon 69 nucleotide T changed to C as a consequence amino acid Serine change to Proline. Conclusions: Sequence variation in viral ORF8 gene is regarded as the primary hotspot for genetic recombination and mutation of spike protein of SARS-CoV-2. Monitoring frequent nucleotide substitutions in ORF8 gene could be extremely helpful in understanding how the virus evolved in the community. Formation of new clades in Erbil city may have impact on vaccination program or the infectivity of the virus. [ABSTRACT FROM AUTHOR]

Enteroviruses (EVs), single‐stranded, positive‐sense RNA viruses, can be classified into four species (A−D), which have previously been linked to a diverse range of disease manifestations and infections affecting the central nervous system. In the Enterovirus species B (EV‐B), Echovirus type 11 (E11) has been observed to occasionally circulate in Taiwan, which was responsible for an epidemic of enterovirus infections in 2018. Here, 48 clinical specimens isolated in 2003, 2004, 2009, and 2018 were collected for the high‐throughput sequencing. Notably, we identified 2018 Taiwanese strains having potential recombinations in the 3D gene, as well as one 2003 strain having a double recombination with E6 and Coxsackievirus B5 in the P2 and P3 regions, respectively. Additionally, one amino acid signature mutated from the Histidine (H) in throat swab specimens to the Tyrosine (Y) in cerebral spinal fluid specimens was detected at position 1496 (or 57) of the genomic coordinate (or 3A gene) to further demonstrate intra‐host evolution in different organs. In conclusion, this study identifies potential intertypic recombination events and an intra‐host signature mutation in E11 strains, isolated during a 2018 neurological disease outbreak in Taiwan, contributing to our understanding of its evolution and pathogenesis. [ABSTRACT FROM AUTHOR]

Conventional breeding involves considerable genetic recombination. To maximize breeding efforts, it is necessary to increase seed yield. This study proposes to improve seed yield by using tetraploid bananas as part of a genetic improvement programme for cooking varieties. To this end, the female fertility (seeding rate and embryo sac maturity) of tetraploid banana plants was characterised. The pollen potential of tetraploids was then assessed at anthesis and at the end of meiosis, as well as during crosses with seminiferous diploids, leading to a phenotypic characterisation of the value of an offspring. This study highlighted the low seed production rate of tetraploid genitors. At best, 7% of seeds are produced per fruit, but 57% of the ovules per fruit have mature embryo sacs. The use of tetraploids as male parents, free from any source of sterility associated with the domestication process, did not generally improve the seed rate, except in the case of FHIA 21 (2930 seeds). Its pollen potential was used in a cross with Banksii 0623 (♀). This resulted in progeny with interesting vigour and phenotypic characteristics. This latter combination has the potential to transfer its "cooking" genetic structure. [ABSTRACT FROM AUTHOR]

Polymers have been used in various industries over the past few decades due to their tremendous applications. Among these, polyhydroxyalkanoates and poly(lactic acid) are easily biodegradable biopolymers derived from bacteria, including recombinant Escherichia coli, Alcaligenes eutrophus, Alcaligenes latus, Azotobacter vinelandii, methylotrophs and Pseudomonas. Conventional petroleum‐derived polymers have become potentially harmful to the environment due to their complex degradation process. The nonbiodegradability of synthetic polymers has become a global issue of concern. There is an urgent need for a substitute to tackle the increasing environmental stress. Microorganisms are small factories for producing different types of polymers during their growth cycle. Various features like biodegradability, biocompatibility, nontoxicity and wide substrate spectrum make such microbial polymers highly reliable. Biopolymers such as alginate, cellulose, cyanophycin, levan, polyhydroxyalkanoates, xanthan, poly(lactic acid) and poly(γ‐glutamic acid) can be obtained from different microorganisms like Aureobasdium pullulans, Acetobacter xylinum, Bacillus thermoamylovorans and Cupriavidusnecator. These are extensively used in various fields like food, medicine, wastewater treatment, biofuel production, packaging and cosmetics. Despite being advantageous in several ways, the biopolymer market still faces several hurdles. This review mainly emphasizes the different types of biopolymers, production by microorganisms and various applications of these biopolymers in different fields. The main drawback limiting the development of these polymers is the high production cost and low efficiency of the microbial strains. Genetic recombination is an efficient technique to enhance the microbial yield and to expand the biopolymer market size. © 2023 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Immune tolerance is maintained in lymphoid organs (LOs). Despite the presence of complex immune cell networks in non-LOs, it is unknown whether self-tolerance is maintained in these tissues. We developed a technique to restrict genetic recombination to regulatory T cells (Tregs) only in skin. Selective depletion of skin Tregs resulted in T cell–mediated inflammation of hair follicles (HFs). Suppression did not rely on CTLA-4, but instead on high-affinity interleukin-2 (IL-2) receptor expression by skin Tregs, functioning exclusively in a cell-extrinsic manner. In a novel model of HF stem cell (HFSC)–driven autoimmunity, we reveal that skin Tregs immunologically protect the HFSC niche. Finally, we used spatial transcriptomics to identify aberrant IL-2 signaling at stromal-HF interfaces in a rare form of human alopecia characterized by HFSC destruction and alopecia areata. Collectively, these results reveal the fundamental biology of Tregs in skin uncoupled from the systemic pool and elucidate a mechanism of self-tolerance. Editor's summary: Regulatory T cells (Tregs) are critical for maintaining peripheral tolerance, but delineation of their tissue-specific functions has been challenging because of limited availability of tools that selectively target tissue Tregs. By developing a technique to induce genetic recombination specifically in murine skin Tregs, Cohen et al. demonstrated that skin Tregs protect the hair follicle from steady-state inflammation. Suppression of hair follicle inflammation required skin Treg expression of the high-affinity interleukin-2 (IL-2) receptor but not CTLA-4. In a mouse model of T cell–mediated autoimmune attack, skin Tregs prevented destruction of hair follicle stem cells. Patients with autoimmune alopecia displayed an enhanced IL-2/STAT5 gene signature around hair follicles, indicating that dysregulation of skin Treg-mediated immunosuppression may contribute to the pathogenesis of autoimmune skin diseases. —Claire Olingy [ABSTRACT FROM AUTHOR]

Group A streptococcal strains potentially acquire new M protein gene types through genetic recombination (emm switching). To detect such variants, we screened 12,596 invasive GAS genomes for strains of differing emm types that shared the same multilocus sequence type (ST). Through this screening we detected a variant consisting of 16 serum opacity factor (SOF)-positive, emm pattern E, emm82 isolates that were ST36, previously only associated with SOF-negative, emm pattern A, emm12. The 16 emm82/ST36 isolates were closely interrelated (pairwise SNP distance of 0–43), and shared the same emm82-containing recombinational fragment. emm82/ST36 isolates carried the sof12 structural gene, however the sof12 indel characteristic of emm12 strains was corrected to confer the SOF-positive phenotype. Five independent emm82/ST36 invasive case isolates comprised two sets of genetically indistinguishable strains. The emm82/ST36 isolates were primarily macrolide resistant (12/16 isolates), displayed at least 4 different core genomic arrangements, and carried 11 different combinations of virulence and resistance determinants. Phylogenetic analysis revealed that emm82/ST36 was within a minor (non-clade 1) portion of ST36 that featured almost all ST36 antibiotic resistance. This work documents emergence of a rapidly diversifying variant that is the first confirmed example of an emm pattern A strain switched to a pattern E strain. [ABSTRACT FROM AUTHOR]

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Summary ChromaX is a Python library that enables the simulation of genetic recombination, genomic estimated breeding value calculations, and selection processes. By utilizing GPU processing, it can perform these simulations up to two orders of magnitude faster than existing tools with standard hardware. This offers breeders and scientists new opportunities to simulate genetic gain and optimize breeding schemes. Availability and implementation The documentation is available at https://chromax.readthedocs.io. The code is available at https://github.com/kora-labs/chromax. [ABSTRACT FROM AUTHOR]

Intervention techniques to restore coral communities have become an important management tool to help recover and rehabilitate damaged reefs. The direct transplantation of healthy coral fragments is the most common method; however, there is controversy in the long-term success, as using coral clones may diminish the genetic diversity of the coral population. Genetic recombination can be achieved when the coral colony produces gametes and eventually reproduces; therefore, it is important to provide evidence that restored colonies produce gametes as their naturally recruited counterparts with similar colony size (age). Natural and restored Pocillopora coral colonies of the same size range (between 40 and 50 cm in diameter) were tagged and sampled during the rainy season to determine gamete maturation. Our results show no differences in the reproductive activity among colonies: natural and restored coral colonies matured gametes from June to October, with a peak in sexually active coral colonies in July. Also, gamete malformation was not observed. During the gamete production period, the area's temperature ranged from 27.9 to 30.02 °C. The results' evidence that coral colonies formed through active restoration contribute not only to the increase in live coral cover as seen in previous studies but potentially contribute in the medium term (>5 years after out-planting) to the production of larvae and local and subsidiary recruitment, since they exhibit the same reproductive patterns as their naturally formed counterparts and no differences in the reproductive activity among coral colonies. Therefore, long-term coral restoration projects contribute to maintaining the live coral cover and the genetic diversity in the region, eventually rehabilitating the coral community. [ABSTRACT FROM AUTHOR]

Background HIV poses significant challenges for vaccine development due to its high genetic mutation and recombination rates. Understanding the distribution of HIV subtypes (clades) across regions and populations is crucial. In this study, a systematic review of the past decade was conducted to characterize HIV-1/HIV-2 subtypes. Methods A comprehensive search was performed in PubMed, EMBASE, and CABI Global Health, yielding 454 studies from 91 countries. Results Globally, circulating recombinant forms (CRFs)/unique recombinant forms (URFs) accounted for 29% of HIV-1 strains, followed by subtype C (23%) and subtype A (17%). Among studies reporting subtype breakdowns in key populations, 62% of HIV infections among men who have sex with men (MSM) and 38% among people who inject drugs (PWIDs) were CRF/URFs. Latin America and the Caribbean exhibited a 25% increase in other CRFs (excluding CRF01_AE or CRF02_AG) prevalence between 2010–2015 and 2016–2021. Conclusions This review underscores the global distribution of HIV subtypes, with an increasing prevalence of CRFs and a lower prevalence of subtype C. Data on HIV-2 were limited. Understanding subtype diversity is crucial for vaccine development, which need to elicit immune responses capable of targeting various subtypes. Further research is needed to enhance our knowledge and address the challenges posed by HIV subtype diversity. [ABSTRACT FROM AUTHOR]

Bacillus thuringiensis Berliner (Bt) and B. cereus sensu stricto Frankland and Frankland are closely related species of aerobic, spore-forming bacteria included in the B. cereus sensu lato group. This group is one of the most studied, but it remains also the most mysterious species of bacteria. Despite more than a century of research on the features of these ubiquitous bacteria, there are a lot of questionable issues related to their taxonomy, resistance to external influences, endophytic existence, their place in multidimensional relationships in the ecosystem, and many others. The review summarizes current data on the mutualistic relationships of Bt and B. cereus bacteria with plants, the structure of the phytomicrobiomes including Bt and B. cereus, and the abilities of plant-associated and endophytic strains to improve plant resistance to various environmental factors and its productivity. Key findings on the possibility of the use of Cry gene promoter for transcription of the target dsRNA and simultaneous release of pore-forming proteins and provocation of RNA-interference in pest organisms allow us to consider this group of microorganisms as unique tools of genetic engineering and biological control. This will open the prospects for the development and direct change of plant microbiomes, and possibly serve as the basis for the regulation of the entire agroecosystem. [ABSTRACT FROM AUTHOR]

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has been widespread globally in pigs and humans for decades. Nasal colonization of LA-MRSA is regarded as an occupational hazard to people who are regularly involved in livestock production. Our previous study suggested pig-to-human transmission caused by LA-MRSA clonal complex (CC) 398, using traditional molecular typing methods. Instead, this study aimed to investigate the zoonotic transmission of LA-MRSA CC398 using whole genome sequencing (WGS) technologies. A total of 63 LA-MRSA isolates were identified and characterized in Thailand. Further, the 16 representatives of LA-MRSA CC9 and CC398, including porcine and worker isolates, were subjected to WGS on the Illumina Miseq platform. Core-genome single nucleotide polymorphism (SNP)-based analyses verify the zoonotic transmission caused by LA-MRSA CC398 in two farms. WGS-based characterization suggests the emergence of a novel staphylococcal cassette chromosome (SCC) mec type, consisting of multiple cassette chromosome recombinase (ccr) gene complexes via genetic recombination. Additionally, the WGS analyses revealed putative multi-resistant plasmids and several cross-resistance genes, conferring resistance against drugs of last resort used in humans such as quinupristin/dalfopristin and linezolid. Significantly, LA-MRSA isolates, in this study, harbored multiple virulence genes that may become a serious threat to an immunosuppressive population, particularly for persons who are in close contact with LA-MRSA carriers. [ABSTRACT FROM AUTHOR]

GII.4 noroviruses have caused the overwhelming majority of norovirus-related gastroenteritis cases during the past two decades. However, a trend towards the emergence of new genotypes and novel GII.4 variants provided the impetus to explore further the changing patterns in norovirus epidemiology during the present study. Genotyping of 60 norovirus strains detected during a period of 33 months (January 2016–October 2018) was performed on the basis of the capsid VP1-coding ORF2 gene sequence. All norovirus strains detected were classified into seven genotypes, six of which belonged to genogroup GII. GII.2 was the dominant genotype till February 2017, whereas GII.4 prevailed thereafter. Most of the GII.4 strains were of the Sydney_2012 variant, whereas five strains could not be classified. Further recombination analysis at the ORF1/ORF2 gene junction revealed that 23 out of 24 strains were recombinant, thereby showcasing the significant role of genetic recombination in norovirus evolution and epidemiology. Continuous genomic surveillance and molecular characterization are essential for tracking norovirus evolution, which could contribute to the elucidation of new aspects of virus–host interactions that potentially affect host morbidity and epidemiology. [ABSTRACT FROM AUTHOR]

Objectives: We conducted molecular characterization, demonstrated the geographical distribution of Zika virus (ZIKV) circulating worldwide from 1947 to 2022 and explored the potential genetic recombination site in the Thailand ZIKV genomes. Methods: We constructed phylogenetic trees based on ZIKV coding sequences (CDS) and determined the geographical distribution of the representative viruses by genetic relationship and timeline. We determined genetic recombination among ZIKV and between ZIKV and other flaviviruses using similarity plot and bootscan analyzes, together with the phylogeny encompassing the CDS and eight subgenomic regions. Results: The phylogenetic trees comprising 717 CDS showed two distinct African and Asian lineages. ZIKV in the African lineage formed two sublineages, and ZIKV in the Asian lineage diversified into the Asian and American sublineages. The 1966 Malaysian isolate was designated the prototype of the Asian sublineage and formed a node of only one member, while the newer viruses formed a distinct node. We detected no genetic recombination in the Thailand ZIKV. Conclusion: Five Thailand isolates discovered in 2006 were the second oldest ZIKV after the Malaysian prototype. Our result suggested two independent routes of ZIKV spread from Southeast Asia to Micronesia in 2007 and French Polynesia in 2013 before further spreading to South American countries.

Coxsackievirus A24 (CV-A24) is a human enterovirus that causes acute flaccid paralysis. However, a Coxsackievirus A24 variant (CV-A24v) is the most common cause of eye infections. The causes of these variable pathogenicity and tissue tropism remain unclear. To elucidate the phylodynamics of CV-A24 and CV-A24v, we analyzed a dataset of 66 strains using Bayesian phylodynamic approach, along with detailed sequence variation and epistatic analyses. Six CV-A24 strains available in GenBank and 60 CV-A24v strains, including 11 Taiwanese strains, were included in this study. The results revealed striking differences between CV-A24 and CV-A24v exhibiting long terminal branches in the phylogenetic tree, respectively. CV-A24v presented distinct ladder-like clustering, indicating immune escape mechanisms. Notably, 10 genetic recombination events in the 3D regions were identified. Furthermore, 11 missense mutation signatures were detected to differentiate CV-A24 and CV-A24v; among these mutations, the F810Y substitution may significantly affect the secondary structure of the GH loop of VP1 and subsequently affect the epitopes of the capsid proteins. In conclusion, this study provides critical insights into the evolutionary dynamics and epidemiological characteristics of CV-A24 and CV-A24v, and highlights the differences in viral evolution and tissue tropism.

Porcine epidemic diarrhea (PED) is a highly contagious disease caused by porcine epidemic diarrhea virus (PEDV), which is characterized by severe diarrhea and vomiting in lactating piglets, resulting in serious economic losses to the pig industry worldwide. Here, a novel variant strain of PEDV (named PEDV/CH/XC/2020) was isolated from the feces of infected piglets, and subjected to genetic variation and recombination analysis based on whole genome sequencing. The results showed that PEDV/CH/XC/2020 belonged to a GIIa subtype variant strain of PEDV, with the genome of 28128 nt in length, which shared only 94% identities with the vaccine strain CV777. Furthermore, multiple amino acid mutations were occurred in the neutralizing antigenic epitopes of COE (499-638 aa) and S1D (636-789 aa) regions of spike protein. It was worth noting that genetic recombination was occurred in the 24010-26546 nt and 27000-27663 nt regions, suggesting that this isolate may arise from genetic recombination with parental strains such as GD-1, CH/ZMDZY and PEDV-CHZ and continuous mutation in epidemic process. In contrast to other mutant strains, the mutation sites 767F, 838L and 1060C of S protein are unique, which might result in the alterations of virulence and immunogenicity of PEDV/CH/XC/2020. These findings of the molecular characteristic of this novel variant strain provided new insights into t